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Live imaging of DORNRÖSCHEN and DORNRÖSCHEN-LIKE promoter activity reveals dynamic changes in cell identity at the microcallus surface of Arabidopsis embryonic suspensions

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Abstract

Key message

Transgenic DRN::erGFP and DRNL::erGFP reporters access the window from explanting Arabidopsis embryos to callus formation and provide evidence for the acquisition of shoot meristem cell fates at the microcalli surface.

Abstract

The DORNRÖSCHEN (DRN) and DORNRÖSCHEN-LIKE (DRNL) genes encode AP2-type transcription factors, which are activated shortly after fertilisation in the zygotic Arabidopsis embryo. We have monitored established transgenic DRN::erGFP and DRNL::erGFP reporter lines using live imaging, for expression in embryonic suspension cultures and our data show that transgenic fluorophore markers are suitable to resolve dynamic changes of cellular identity at the surface of microcalli and enable fluorescence-activated cell sorting. Although DRN::erGFP and DRNL::erGFP are both activated in surface cells, their promoter activity marks different cell identities based on real-time PCR experiments and whole transcriptome microarray data. These transcriptome analyses provide no evidence for the maintenance of embryogenic identity under callus-inducing high-auxin tissue culture conditions but are compatible with the acquisition of shoot meristem cell fates at the surface of suspension calli.

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Abbreviations

CaMV:

Cauliflower mosaic virus

CIM:

Callus-inducing medium

FACS:

Fluorescence-activated cell sorting

SAM:

Shoot apical meristem

SE:

Somatic embryogenesis

SEIM:

Somatic embryogenesis inducing medium

SIM:

Shoot-inducing medium

GFP:

Green fluorescent protein

erGFP:

Endoplasmic reticulum targeted GFP version

DRN = ESR1 :

DORNRÖSCHEN or ENHANCER OF SHOOT REGENERATION1

DRNL = ESR2 :

DORNRÖSCHEN-LIKE or ENHANCER OF SHOOT REGENERATION2

WOX :

WUSCHEL-related homeobox

WUS :

WUSCHEL

STM :

SHOOT MERISTEMLESS

ARF :

Auxin response factor

IAA :

Indoleacetic acid-induced protein

ARR :

Arabidopsis response regulator

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Acknowledgments

This work was funded by the Deutsche Forschungsgemeinschaft via SFB 572.

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Authors and Affiliations

  1. Institut für Entwicklungsbiologie, Biozentrum, Universität zu Köln, Zülpicher Str. 47b, 50674, Köln, Germany

    M. Cole, B. Jacobs, J. W. Chandler & W. Werr

  2. Unité de Recherche en Génomique Végétale (URGV), UMR INRA 1165 - Université d’Evry Val d’Essonne - ERL CNRS 8196, 2 rue G. Crémieux, CP 5708, 91057, Evry Cedex, France

    L. Soubigou-Taconnat & S. Balzergue

  3. Institut de Recherche en Horticulture et Semences, UMR 1345 (INRA; Université d’Angers) Centre Angers-Nantes, 42 rue Georges Morel, BP 60057, 49071, Beaucouzé cedex, France

    J. P. Renou

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  1. M. Cole
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  2. B. Jacobs
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  3. L. Soubigou-Taconnat
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  4. S. Balzergue
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Correspondence to W. Werr.

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Communicated by R. Schmidt.

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Cole, M., Jacobs, B., Soubigou-Taconnat, L. et al. Live imaging of DORNRÖSCHEN and DORNRÖSCHEN-LIKE promoter activity reveals dynamic changes in cell identity at the microcallus surface of Arabidopsis embryonic suspensions. Plant Cell Rep 32, 45–59 (2013). https://doi.org/10.1007/s00299-012-1339-4

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  • DOI: https://doi.org/10.1007/s00299-012-1339-4

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